Hepatitis C virus (HCV) specific sequences are demonstrable in the DNA fraction of peripheral blood mononuclear cells from healthy, anti-HCV antibody-negative individuals and cell lines of human origin.
No convincing support has been provided so far for the existence of extrahepatic hepatitis C virus particles that should correspond to the sometimes extremely high concentration of 'HCV-RNA' in serum or plasma. If a naturally occurring HCV-specific DNA were to be found, a concept for at least some phenomena in terms of the pathophysiology of HCV should become conceivable. DNA was extracted from peripheral blood mononuclear cells of eleven healthy, anti-HCV-negative individuals, including five long term blood donors, and cells from different cell lines. DNA was subjected to nested polymerase chain reaction omitting a reverse transcriptase step with primers of the 5'NC as well as part of the core region of HCV. Direct polymerase chain reaction, i.e. without a reverse transcriptase step, revealed HCV-specific sequences in the DNA fraction of peripheral blood mononuclear cells of different origin: healthy anti-HCV negative individuals, furthermore in HeLa and MT2 cells. The fragments found were of expected length as well as of shorter and of longer than expected length with respect to the sequence of the HCV genome framed by the primers applied. The results derived from additional hybridization, restriction endonuclase analysis, and sequencing demonstrated HCV-specific sequences in the expected fragments with both a high degree of homology and deletions, respectively, substitutions, as compared to a prototype strain. However, the longer than expected fragments also contained sequences not specific for HCV.
Dennin
R H
RH
Institut für Medizinische Mikrobiologie und Hygiene, Medizinische Universität zu Lübeck, Germany.
Chen
Z
Z
D001483:Base Sequence; D002460:Cell Line; D004247:DNA / Q000097:blood*; D017931:DNA Primers; D016174:Hepacivirus / Q000276:immunology / Q000378:metabolism*; D018937:Hepatitis C Antibodies / Q000097:blood* / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D009000:Monocytes / Q000821:virology*; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D012367:RNA, Viral / Q000737:chemistry; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
D017931:DNA Primers; D018937:Hepatitis C Antibodies; D012367:RNA, Viral; D004247:DNA